This is a quick summary of Dr.Frye's July 2012 paper, "Biomarkers of Energy Metabolism in ASD Children." Caveat - this summary is by a layman, might have errors etc.

1. A LAYMAN’S SUMMARY OF
“Biomarkers of Abnormal Energy Metabolism In Children
with ASD”, Richard Frye, MD PhD, NAJMS July 2012
C AV E AT: I a m a p a r e n t o f a n A S D c h i l d a n d I h a v e n o
m e d i c a l t r a i n i n g . I h a v e s u m m a r i z e d D r. F r y e ’s p a p e r
( a v a i l a b l e a t w w w. n a j m s . n e t / v 5 i 3 p 1 4 1 w / ) to t h e
best of my ability in the hope that it may help busy
parents access this important material. There may
be errors of understanding here. If you have
comments or corrections, e-mail –
p a r e n t s . g r o u p . M A P S . f o r. a u t i s m @ g m a i l . c o m

2. FIRST, A WORD ABOUT THIS FILE
NOTES:
 You are seeing this file because you are a member of the Facebook Group “MAPS for
Autism – Parents Group” or because this file was re-posted by one of the members on
another group.
 MAPS is an organization whose mission is to provide biomedical physicians with
training and long-term support, to ensure the quality and consistency of care for
children with autism and related chronic conditions. For more info on MAPS
Phsyicians, see www.medmaps.org
 MAPS for Autism – Parents Group is a parents-only group for parents who see MAPS
physicians or are interested in the treatments. This is an informal parents-only group
that is NOT endorsed by the MAPS organization in any way, shape or form.
 If you are interested in joining this group, search Facebook for the group name and ask
to join; if you have any problems with this, e-mail the Group Admin at
maps.for.autism.parents.group@gmail.com

3. AS A PARENT, WHY SHOULD YOU CARE ABOUT
THIS STUDY?
 The following is my opinion as a parent
 Prior studies in ASD kids looked at some biomarkers to
figure out, “Do ASD kids have mitochondrial disease”?
 This study goes further.
 It looks at a variety of biomarkers in ASD kids.
 It also links abnormalities in biomarkers to physiologic
abnormalities in autism.

4. WHAT IS THE PURPOSE OF THIS STUDY?
Before I answer that, I’ll first share some info about the research on mito dysfunction in ASD kids -
 According to a recent study by Frye & Rossignol, about 5% of ASD kids have classic markers for
mitochondrial disease
 These children have clinical symptoms different from the general ASD population. This sub-group
of kids is called the autism/mitochondrial disease (ASD/MD) group
 According to various other studies, about 30-80% of ASD kids have impaired mitochondrial
function
 Now, 5% - 80% is quite a range! Why such a wide variance? This is because the studies all used
different biomarkers to study mitochondrial function in ASD kids
So back to the question … what is the purpose of this study?
 This study tries to address some of the limitations of earlier studies.
 It looks at a large number of biomarkers in a large sample of ASD kids (133 kids.)
 It looks to answer the questions – how many ASD kids actually have impaired mitochondrial
function? What does this mean in terms of other markers of mitochondrial function?

5. WHAT BIOMARKERS WERE LOOKED AT?
The study specifically looked at these biomarkers in a
morning blood sample with overnight fasting:
 Plasma lactate
 Plasma alanine
 Alanine/Lysine ratio
 Creatine Kinase
 AST level (a measure of liver function)
 Plasma acylcarnitines
If there was an abnormal value, the testing was
repeated.

6. DIAGNOSES AND DEVELOPMENTAL ISSUES IN THE
KIDS IN THE STUDY
Each child in the study had one of the following clinical diagnoses -
 Classic autistic disorder (AD) with no motor delay
 PDD-NOS with no motor delay
 AD with motor delay
 PDD-NOS with motor delay
 Isolated speech delay
 ADHD (with hyperactivity)
 ADHD (without hyperactivity)
The study also looked at clinical characteristics like whether
the child had epilepsy or a developmental regression.

7. STUDY FINDINGS

8. STUDY FINDINGS
Over 30% of the children in the sample of 133 were found to have metabolic
abnormalities. Here is a summary. There are lots more tables of results, look at
the Frye paper for more data.
Biomarker % of kids with
abnormalities
Lactate 16.9%
Alanine 1.7%
Alanine/Lysine Ratio 15.9%
Acylcarnitines 23.8%
AST 10.1%
CK 6.8%

9. THE STUDY FOUND FOUR DISTINCT SUB-GROUPS
Of the children with metabolic abnormalities, there were four
distinct sub-groups -
 Sub-group 1 – Abnormally elevated lactate
 Sub-group 2 – Abnormally elevated AST
 Sub-group 3 – Abnormally elevated alanine/lysine ratio
 Sub-group 4 – Abnormal elevations in multiple acylcarnitines
The sub-groups had some over-lap i.e. some kids could be in
more than one group at once. But there wasn’t a whole lot
of overlap. Let’s take a look at these subgroups 

10. THE STUDY FOUND FOUR DISTINCT SUBGROUPS
TWO SUB-GROUPS MAY HAVE TWO SUB-GROUPS MAY HAVE
MITOCHONDRIAL DYSFUNCTION OTHER ISSUES, NOT MITO
Sub-group 1 – Abnormally elevated lactate Sub-group 2 – Abnormally elevated AST
 This sub-group may indeed have  ASD children with elevated AST values
mitochondrial dysfunction may have oxidative stress rather than
 There is no genetic abnormality common to mitochondrial dysfunction.
all children in the group
Sub-group 3 – Abnormally elevated alanine-to -
lysine ratio Sub-group 4 – Abnormal elevations in
 ASD children with abnormally elevated multiple acylcarnitines
alanine/lysine ratio may indeed have  ASD children with this pattern of elevated
mitochondrial dysfunction associated acyl carntines may not have
with a Complex I deficiency. mitochondrial dysfunction.
 This is not due to a genetic abnormality  Data from an animal model suggests that
common to all children in the group. these metabolic abnormalities may be
associated with propionic acid created by
a bacteria species called clostridia.

11. MORE DETAILS ON METABOLIC
ABNORMALITIES IN EACH SUB-
GROUP

12. SUB-GROUP 1 – ELEVATED LACTATE
FIGURE: Metabolic biomarkers
which demonstrate significant
differences between a
subgroup with consistently
elevated lactic acid and a
control group of ASD children
without metabolic
abnormalities.
Children with abnormally elevated lactate had -
 Elevated urine 2-methyl-3-hydroxybutyric acid which may be due to an
inefficient citric acid cycle
 Higher values for ammonia than controls
 A higher rate of motor delays
CONCLUSION: This sub-group of ASD children may indeed have
mitochondrial dysfunction.

13. SUB-GROUP 2 – ABNORMALLY ELEVATED AST VALUES
 AST is a marker for liver function FIGURE: Metabolic biomarkers which demonstrate significant
differences between a subgroup of children with consistently
 Compared to ASD controls, those with elevated AST and a control group of ASD children without
highly elevated AST also had lower urine metabolic abnormalities.
5-oxoproline (also known as
pyroglutamate)
 Pyroglutamate is a metabolite of the
gamma-glutamyl cycle which is involved
in glutathione utilization and recovery
 Low urine 5-oxoproline may mean
glutathione depletion, which reduces the
liver’s ability to protect itself against
oxidative stress and neutralize toxins
 This could cause liver dysfunction
resulting in increased AST
CONCLUSION:
ASD children with elevated AST values may have oxidative stress
rather than mitochondrial disease.

14. SUB-GROUP 3 – ABNORMALLY ELEVATED
ALANINE/LYSINE RATIO
 Compared to controls, ASD children
FIGURE: Metabolic biomarkers which demonstrate significant
with elevated alanine/lysine ratio also differences between a subgroup of children with consistently
had abnormally elevated alanine and elevated alanine-to-lysine ratio and a control group of children
without metabolic abnormalities.
urine pyruvate
 Lactate was not abnormally elevated
like Sub Group 1, but it was still higher
than controls
 These children had a higher rate of
epilepsy
 There were no genetic abnormalities found
that were common to all children in the
group
 These metabolic abnormalities may be
associated with a mito Complex I deficiency
CONCLUSION: ASD children with abnormally elevated alanine/lysine ratio may indeed
have mitochondrial dysfunction, which is not due to a genetic abnormality common to
all children in the group; this may be associated with a Complex I deficiency.

15. SUB-GROUP 4 – CONSISTENT ELEVATIONS IN
ACYLCARNITINES
ASD children with consistent abnormalities in
acylcarnitines were found to have - FIGURE: Metabolic biomarkers which demonstrate significant
 Higher C5OH, C12, C14, C14:OH and C16 differences between a subgroup of children with consistent
acylcarnitines – i.e. carnitines associated with elevations in multiple acyl-carnitines and a control group of
children without metabolic abnormalities.
short & long chain but not medium-chain fatty
acids; this pattern of acylcarnitine elevations is
not consistent with any known fatty oxidation
disorder
 Higher urine 3-OH-3-methylglutaric acid, which
suggests citric acid cycle abnormalities
 This pattern is consistent with abnormalities
seen in a rodent model when rodents were
injected with propionic acid
 This sub-group of children has a high rate of
regression
 Propionic acid can be produced by Clostridia , a
bacterial species seen in kids with regressive ASD
CONCLUSION: ASD children with this pattern of elevated acylcarntines may not have mitochondrial
dysfunction. Data from an animal model suggests that these metabolic abnormalities may be
associated with propionic acid created by a bacteria species called clostridia.